Abstract
Due to the increase in the incidence of Clostridium difficile associated diseases at a tertiary care center in Lebanon, this study was undertaken to determine the prevalent C. difficile toxinotypes. The immunocard method was used to test for toxins A and B in 88 collected stool samples, followed with API 20A to confirm for C. difficile. PCR amplification of the triose phosphate isomerase (tpi) gene, the toxin encoding genes tcdA, and tcdB, followed by toxinotyping, were performed on recovered isolates and stool specimens. Out of the 88 stool samples obtained, 30 (65.2%) were Immunocard positive, culture and or tpi positive for C. difficile. Of the 30 isolates, 4 were PCR negative for the tcdA and tcdB genes (A-B-), and 26 were PCR positive for the tcdA and / or tcdB genes with 4 being A+B+, 1 A+B-, and 21 A-B+. The results of toxinotyping showed that 2 isolates belonged to toxinotype 0, 4 to toxinotype XI, 2 to toxinotype XII, 1 to toxinotype XVI, 1(A+B-) and twenty (A-B+) designated as toxinotype 0-like. C. difficile was detected in 65.2% of patients' stools with prevalence of toxinotype 0-like. Identification of toxinotypes of C. difficile is important to determine the virulence potential of strains and control their spread.
Highlights
Due to the increase in the incidence of Clostridium difficile associated diseases at a tertiary care center in Lebanon, this study was undertaken to determine the prevalent C. difficile toxinotypes
Pseudomembranous colitis is an inflammation of the colon often caused by the bacterium C. difficile, and is directly associated with simultaneous prolonged hospitalization of infected people who will be put on antibiotics
Toxinotyping of the 24 isolates revealed 2 (7.69%) to be of toxinotype 0, 2 (7.69%) of toxinotype XII, 1 (3.84%) of toxinotype XVI, 1 (3.84%) of a new toxinotype having a phenotype (A+B-) where the restriction patterns of A3 is of type 1 and 20 (76.92%) were of a new toxinotype having a phenotype of (A-B+) where the restriction pattern of B1 is of type 1, this group was designated as toxinotype 0-like (Figures 1 and 2)
Summary
Due to the increase in the incidence of Clostridium difficile associated diseases at a tertiary care center in Lebanon, this study was undertaken to determine the prevalent C. difficile toxinotypes. Methodology: The immunocard method was used to test for toxins A and B in 88 collected stool samples, followed with API 20A to confirm for C. difficile. PCR amplification of the triose phosphate isomerase (tpi) gene, the toxin encoding genes tcdA, and tcdB, followed by toxinotyping, were performed on recovered isolates and stool specimens. Results: Out of the 88 stool samples obtained, 30 (65.2%) were Immunocard positive, culture and or tpi positive for C. difficile. C. difficile was detected in 65.2% of patients' stools with prevalence of toxinotype 0-like. C. difficile can cause toxin-mediated diseases including postantibiotic diarrhea and extending to severe pseudomembranous colitis (PMC). Pseudomembranous colitis is an inflammation of the colon often caused by the bacterium C. difficile, and is directly associated with simultaneous prolonged hospitalization of infected people who will be put on antibiotics. Two main virulence factors are involved, toxins TcdA and TcdB encoded by the toxin genes, tcdA and tcdB on the PaLoc (pathogenicity locus) region [4].
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